Part:BBa_P0140

Tianjin’s 2021 characterization TetR BBa_P0140

New information of TetR promoter leakage

Background

In order to better apply the TetR promoter in the project, we characterized the leakage amount of the TetR promoter. We constructed a carotene plasmid with TetR promoter, and transformed it into yeasts, then cultured the yeasts without AHT inducer. By observing the color of the colony, the leakage of TetR promoter can be roughly determined.

Result

Our experimental results show that the TetR promoter is very easy to leak, and the expression amount of the circuit is different and difficult to test. Therefore, the results clearly indicated that in strict experiments, using the TetR promoter alone will affect the experimental results. <p> <img style=”margin-left:10%” width=”80%” src="">

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   <img alt="" src="" width="400" height="313" class="thumbimage" srcset="/wiki/images/d/dd/Wro_drop.png 1.5x, /wiki/images/d/dd/Wro_drop.png 2x"></a>  

   Figure 1. Drop test analysis of <i>S. cerevisiae EBY.VW4000 transformed with pRS426 plasmid containing YALI0C06424g, YALI08943g and YALI0F19184g hexose transporters from Y. lipolytica under the control of ADH1 (pA) or TEF (pT) promoter.  </i>

Design

<img style=”margin-left:10%” width=”80%” src=" " >

Results